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Laboratory Culture of Musca, Fannia, and Stomoxys

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Laboratory Culture of Musca, Fannia, and Stomoxys Bull. Org. mond. Santw Bull. Wid. Hlth Org. 1964, 31, 539-544 Laboratory Culture of Musca, Fannia, and Stomoxys H. F. SCHOOF 1 In spite of great improvements since 1940, diffi- Larvae culties are still encountered in rearing the large Various media have been used for rearing: horse numbers of flies of uniform quality required by manure (Glaser, 1924), horse manure and additives modern entomological research. These difficulties (Glaser, 1927; Hockenyos, 1931), and cow manure partly account for variations in techniques, rearing (Feldman-Muhsam, 1944). Richardson (1932) pro- media and equipment. posed the alfalfa-meal/wheat-bran/yeast formula that was the forerunner of the current CSMA mash REARING MUSCA (i.e., alfalfa meal 27%, wheat bran 33%, Brewer's dried grain 40%) and of the CSMA medium (Soap Musca domestica, the housefly, because of its Blue Book, 1960), which consists of 340 g of the ease of handling and ability to propagate in widely CSMA mash mixed with 750 ml of aqueous sus- varying conditions, is probably the species most pension that contains 15 g of moist yeast cake and commonly used for evaluation of insecticides. This 10 ml of diamalt. As usually occurs, this standard fact, together with the tendency to use different preparation 2 has been modified by various labo- methods in the various laboratories, either through ratories according to their own needs or desires choice or because of factors such as space, money, (WHO Expert Committee on Insecticides, 1961; and volume of insects required, has led to the use of Gahan, 1963; Basden, 1947). The YMA medium a number of slightly different techniques. (WHO Expert Committee on Insecticides, 1961; Rockstein, 1957) consists of dried yeast, 100 g; Eggs full-cream dried milk, 100 g; agar, 20 g; and boiling Housefly eggs are deposited in clusters or masses water, 1 litre. Larvae have been reared on cotton- upon substrates placed in the colony cages for that wool pads soaked with diluted milk (milk: water, purpose, e.g., a cellulose cotton pad 3 inches (7.5 cm) 3: 1) (Hafez, 1948) and on fermenting dog-biscuits in diameter and 1 inch (2.5 cm) thick and soaked in (Frings, 1948). milk and honey-water,2 dental cotton-wool rolls For Musca autumnalis, Fales (1963) found cow impregnated with a weak solution of milk powder manure to be the only medium that sustained the and dried yeast (Sawicki and Holbrook, 1961), larvae to maturity. In India, coconut poonac is an crimpled whole oats (Eagleson, 1937), filter-paper excellent substrate for M. d. vicina (Tharumarajah & soaked with milk and a few drops of ammonia Thevasagayam, 1961); in Japan a mixture of (Okara) (Fisher & Morrison, 1949), fermenting dog-biscuits tofu, 50 g, rice bran, 5 g, and yeast powder, 0.5 g (Frings, 1948), and numerous other materials (Soap is used (Nagasawa & Hashizume, 1955). Other Blue Book, 1960). After 18-24 hours the eggs hatch media for M. domestica have included crimpled when the pads are moistened with or immersed in whole oats, syrup and yeast (Eagleson, 1937), water.2 Egg quantity can be calculated volumetri- guinea-pig meal (Hampton, 1952), coffee-grounds cally, 500 (West, 1951) to 750 (Smith and Harrison, and cheese (Hase, 1935) and tissue-paper, milk 1951) per 0.1 ml or weighed (1100 newly-hatched powder and yeast (Spiller, 1963). Media are hand- larvae = 90 mg). mixed or mechanically mixed; the main point is to mix thoroughly to ensure uniform moisture distri- bution. The finished medium 2 is 24 hours 1 Assistant Chief, Biology/Chemistry Section, Technology prepared Branch, Communicable Disease Center, Public Health in advance of use and 2 pounds (900 g) are placed Service, US Department of Health, Education, and Welfare, in a plastic bag that is inserted in a 1-US-gallon Savannah, Ga., USA. ' Unpublished memoranda of the Communicable Disease (3.7-litre) battery jar. Packing the medium is Center Laboratories, Savannah, Ga. avoided. Newly-hatched larvae are placed on the 1507 -539- 540 H. F. SCHOOF medium and 3-4 days later a 3-inch (7.5-cm) layer supply of water (Gahan, 1963); or agar, banana, of sand is added.' Other seeding rates are 5000 eggs sugar, skimmed milk, formol, gelatin and water in 10 litres of medium (Sawicki & Holbrook, 1961) (Eagleson, 1943). Milk in liquid or dry form is and 2000 eggs per 75 g of a dry mix of milk powder, essential to flies used for egg production but not to absorbent tissue-paper and yeast with 2.1 ml water those used for test purposes. per g of mix (Spiller, 1963). For CSMA media, temperatures up to 130°F (54°C) are considered Adult colonies allowable. Odour from fermentation can be mini- In susceptible colonies 3000 pupae are placed in a mized by continually ventilating the rearing room. cage 14.5 x 16.5 x 19 inches (about 36 x 41 x 46.5 cm) and provided with sugar cubes, powdered milk, and Pupae water. An oviposition pad is put in the cage over- At 80°F (260C) and 70 % relative humidity, larvae night. Cages which provide less than 1 cubic inch begin to pupate in the upper portion of the medium of space per fly reduce the fertility and longevity -i.e., sand, medium (Soap Blue Book, 1960), of adults. At 80°F (26°C) Rockstein & Lieberman vermiculite, sawdust (Sawicki & Holbrook, 1961)- (1959) reported males to have a shorter life-span 4-5 days after the eggs are set. Pupation is usually than females (17 days, 3, 30 days ?). Normally, complete by day 7. The sand is then poured from houseflies begin oviposition on the third or fourth the jar through a sieve to remove the pupae.1 In the days after emergence, with the majority of eggs CSMA technique, the upper portion of the medium deposited within the first 10 days of oviposition is removed, poured into a tray and allowed to dry (Wilkes et al., 1948; Buei, 1959). Adults of M. in the air. autumnalis are confined in cage 1Ox 1Ox 10 inches Pupae are separated by directing an air current (25 x 25 x 25 cm) and fed skimmed milk: sugar against the medium-pupae mixture. The upper, (2: 1 v/v) and a small amount of diamalt solution puparia-laden, stratum also may be put in water, containing brain-heart infusion and pollen (Fales, where the medium sinks and the pupae float (Gahan, 1963). Citrated beef blood and cow manure are 1963). Born (1954) found sand to be a deterrent to also provided. Female M. autumnalis begin egg mould growth when a 1-inch (2.5-cm) layer was deposition at 6 days of age. Fresh cow manure added to the medium 48 hours after it had been serves as the egg-laying medium. M. autumnalis seeded. Puparia can be calculated in number either does not oviposit readily except in sunlight or high by weight or by volume. Normally, pupae have an artificial illumination. average weight of 18-22 mg and average 25 per ml. Life-cycle FACTORS INFLUENCING FLY DEVELOPMENT At 80°F (26°C), the time sequences are: eggs Temperature, humidity and light hatch, 16-24 hours; larvae mature, days 3-4; pupae present, days 4-5; adults emerge, days 8-10; ovi- Generally, the period of development for each position begins, day 12 or 13. stage decreases as the temperature rises. At 74°F (24°C) the eggs of M. domestica required 17 hours Adult food and care before hatching whereas at 99°F (37°C) the period Each holding cage is supplied daily with one of was 7.6 hours (Davidson, 1944). Eggs of M. d. the following: a solution of 5 % spray-dried non-fat vicina hatched in 6 hours at 95°-100°F (35°C-38°C) milk with solids and 2% sugar (Soap Blue Book, and in 11 hours at 77°-86°F (25°C-30°C) (Hafez, 1960); dry powdered milk (Sawicki & Holbrook, 1941). Adult M. domestica are active from 44°F 1961); granulated sugar, liquid milk and 10% to 1 10°F (7°C-43°C) with the optimum at 92°F- sucrose; sugar cubes and water; 1 solid sugar and a 93°F (33°C-33.50C) (Nieschulz, 1935). At 60°F mixture of equal parts of cow's milk and water (15.5°C) adults survived up to 91 days (Dove, 1916). plus 1: 2000 parts of formol as a preservative; a dry The effect of humidity is usually combined with mixture containing 6 parts granulated sugar, 6 parts temperature. However, excessive moisture is detri- powdered milk, 1 part powdered egg and a separate mental to larval development. The significance of light in rearing procedures for M. domestica appears but with M. 1 Unpublished memoranda of the Communicable Disease minor, autumnalis strong artificial light Center Laboratories, Savannah, Ga. or sunlight is necessary for oviposition (Fales, 1963). LABORATORY CULTURE OF MUSCA, FANNIA AND STOMOXYS 541 Crowding length of adult stage, egg production or egg viability Too many larvae for a given supply of food results (Pimentel et al., 1951). No differences were noted in increased larval mortality and smaller adults. in six susceptible and two DDT-resistant strains in Adult crowding causes excessive activity, com- regard to egg viability, length of larval cycle or adult petition, decreased longevity, and greater mortality. production rate (Babers et al., 1953) but in Denmark An increase in the diameter of the rearing container the larvae of a DDT-resistant strain were found to from 4 cm to 7 to 10 cm resulted in the period develop faster than those of a susceptible one between oviposition and the emergence of female (B0ggild & Keiding, 1958).
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